The objective of this proposal is to validate potential biomarkers of excessive alcohol exposure discovered in the Phase 1 portion of this work (1R43AA016233-01). This will entail the development of pipettor tip based immuno-affinity mass spectrometry (AMS) devices for the routine and repetitive high throughput analysis of the biomarkers in question. Resultant devices will allow for both quantitative and qualitative differentiation of the markers of interest. With the successful development of immuno-AMS devices for the biomarkers in question, they will be used to screen clinical samples collected by our clinical collaborators (Aurora Psychiatric Hospital, Wauwatosa, WI) in order to validate their performance. Final metrics for the evaluation of the biomarker assays developed will be a clinical sensitivity and specificity of 85%, with coefficients of variation of <15%. PUBLIC HEALTH RELEVANCE: Alcohol abuse has a significant impact on our society, yet there are no accurate biomarker tests for prognostic, diagnostic or treatment efficacy assessment currently available. This application is to validate new biomarkers of excessive alcohol exposure using a novel in pipettor tip immuno-affinity mass spectrometry based assay. Clearly, accurate and specific biomolecular assays are of great importance, positively impacting the associated health, financial and emotional repercussions of alcohol abuse.